Chapter-5

Experimental
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5. MATERIALS AND EQUIPMENTS

5.1 Materials
 Chemicals: : Lornoxicam, Flurbiprofen, Aceclofenac, Piroxicam, pH 7.4
buffer, potassium dihydrogen phosphate, sodium hydroxide, disodium
hydrogen phosphate, sodium hydroxide, n-octanol, Pluronic F-127, lecithin,
isopropyl myristate, Sodium sorbate, Sodium Benzoate, distilled water,
ethanol, methanol, and acetone.

 Glassware: Beaker, volumetric flask, glass rod, conical flask, funnel, pipette,
measuring cylinder, separating funnel, slides, test tubes, capillary.

 Equipment: UV double beam, stage micrometer, microscope, ocular lense,

Thiels melting point apparatus, hot air oven, pH meter, electronic balance,
dessicator.

5.2 Preformulation studies

1. Determination of solubility
a. Qualitative solubility
Qualitative solubility analysis of drugs were done by dissolving 5 mg of drug
in 5 ml of distilled water and different solvents such as HCl (0.1N), NaOH (0.05
N), Saline phosphate buffer (pH 7.4), Phosphate buffer(pH 9), Phosphate buffer
(pH 4), phosphate buffer (pH 2), ethanol, methanol, acetone and chloroform were
used to determine the solubility of drug.

b. Quantitative Solubility
Quantitative solubility analysis of drugs were done by 5 ml each solvent and
drug in gm(s) into the solvent till saturation of solvent. Different solvents were
used for the solubility determination like distilled water, Saline phosphate buffer
(pH 7), Phosphate buffer(pH 3.6), HCl (0.1N) and NaOH (0.05N). This is done to
determine the capacity of the solvent for dissolving the drug in it. The
concentration of drug is measured by UV spectrophotometer at 376 nm.

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2. Partition Coefficient
The 10 mg of drug was dissolved in 10 ml of distilled water and 10 ml of
carbon tetra chloride in separating funnel & shaken well for 3-4 hours and then
allowed to stand for at least 1 hour for phase separation. After that the water phases
were separated out and the concentration of drug was measured
spectrophotometrically after suitable dilution at 376 nm.

Po/w = Coil/Cwater

3. Melting Point
In this the presealed capillary was filled by the small amount of drug and the
capillary was placed in Melting Point Apparatus.The temperature when the drug starts
to melt and the temperature when drug complete melting was noted.

4. Particle Size

I) Calibration of eyepiece: Use standard stage micrometer to calibrate the eyepiece

micrometer and calculate for the least count (1 eye piece division)
Least Count = No. Of Stage Division / No. Of Eye Piece Div x 10.

II) Mounting of The sample: Transfer a small portion of the given sample on clean
slide and disperse it uniformly and place the slide on the stage of microscope.

III) Measurement of particle size: Focus the slide in low magnification (10x).
observe the particles than shift to high power (45x) and focus the slide. Measure the
size of each particle in terms of eyepiece divisions,a total of 100 particles should be
considered,tabulate the particles in terms of division of eyepiece and no. of particles
(frequency) obtained above,classify the diameter into size ranges and average
frequency of particles in terms of no. distribution.

5. Drug- excipient interaction study

A small amount of drug substance with excipients (like plronic F-127,
Lecithin, Sodium sorbate, Sodium benzoate, Isopropyl myristate, PEG 400,

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triethanolamine, carbopol 934, oleic acid, ethanol and propylene glycol) that is,
physical mixture of the drug and excipients (in 1:1 ratio were prepared to have
maximum likelihood interaction) were placed in a vial, and rubber stopper was placed
on the vial and sealed properly. A storage period of 2 weeks at 60°C, and the same
sample was retained for 2 months at 40°C. After storage the sample were observed
physically for liquefaction, caking, odour or gas formation, & discolouration.

6. Determination of λ max

A. Lornoxicam
The identification of drug was done by UV spectrophotometric method
reported by Nemutlu et al (2005). The small amount of drug is dissolved in 0.05 N
NaOH and scanned in UV range 200-600 nm in UV Double beam spectrophotometer.
The highest peak was determined, which is the λmax for the Lornoxicam .The spectral
data from this scan was used for the preparation of standard curve of Lornoxicam.

B. Flurbiprofen
100 mg of Flurbiprofen was dissolved in 100ml of ethanol. One ml of the
prepared stock solution was further diluted to 100 ml and finally scanned for
maximum absorbance using U.V. spectrophotometer in the range from 230 to 360 nm.
Average of triplicate readings was taken.

C. Aceclofenac
Aceclofenac was dissolved in PBS to obtain a 1000mcg/mL solution. This
solution was subjected to scanning between 200 – 400 nm and absorption maximum
was determined. The effect of dilution on absorption maxima was studied by diluting
the above solution to 20mcg/mL and scanned from 200 – 400nm.

D. Piroxicam
The identification of piroxicam done by UV spectrophotometer method. The small
amount of drug dissolve in 0.1N Hydrochloric acid and scanned in UV. The highest
peak is λmax for the piroxicam. From the spectra 333 nm ( λmax of piroxicam) was
obtained.

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2) Preparation of standard curve

A. Lornoxicam
a. Standard curve of lornoxicam in 0.05 N NaOH
Standard stock solution of Lornoxicam was prepared by dissolving 100 mg
drug in 100 ml 0.05 N NaOH (i.e.1000μg/ml). Aliquot of this solution are further
prepared by taking 10 ml of above solution and diluting it upto 100 ml (i.e. 100
µg/ml). From the above solution further dilutions are prepared in range of 5-35 µg/ml.
Absorbances were taken on UV spectrophotometer at 376 nm against 0.05 N NaOH as
a blank. From these absorbance’s, the standard curve is plotted. Standard curve
equation and regression value is obtained. The absorptivity coefficient of drug at
desired wavelengths was determined.

b. Standard curve of lornoxicam in 7.4 PBS

Standard stock solution of Lornoxicam was prepared by dissolving 10 mg drug
in 10 ml 7.4 PBS (i.e.1000μg/ml) containing 12mg of Tromethamine. This solution
was then sonicated till complete dissolution of drug. Aliquot of this solution are
further prepared by taking 10 ml of above solution and diluting it upto 100 ml (i.e. 100
µg/ml). From the above solution further dilutions are prepared in range of 4-24 µg/ml.
Absorbances were taken in UV spectrophotometer at 376 nm against 7.4 PBS as a
blank.From these absorbance’s, the standard curve is plotted. Standard curve equation
and regression value is obtained. The absorptivity coefficient of drug at desired
wavelengths was determined.

B. Flurbiprofen
a. Standard curve of Flurbiprofen in ethanol
100 mg of Flurbiprofen was accurately weighed and dissolved in ethanol in a
100 ml volumetric flask and the volume was made upto the mark with ethanol. The
above prepared solution of Flurbiprofen was subsequently diluted with ethanol to get
2, 4, 6, 8, 10, 12 μg per ml of the final solution. Then the absorbance was measured by
spectrophotometer at 248nm using ethanol as blank. Average of triplicate readings was
taken.
b. Standard curve of flurbiprofen in 7.4 PBS
Standard stock solution of Flurbiprofen was prepared by dissolving 10 mg drug
in 10 ml 7.4 PBS (i.e.1000μg/ml). Aliquot of 10 ml solution are further taking 10 ml of

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diluted upto 100 ml (i.e. 100 µg/ml). From the above solution are prepared in range of
10-90 µg/ml. Absorbances were taken on UV spectrophotometer at 248 nm against 7.4
PBS as a blank. From these absorbance’s, the standard curve is plotted. Standard
curve equation and regression value is obtained.

C. Aceclofenac
a. Standard curve of Aceclofenac in PBS
A stock solution containing 1 mg/mL of pure drug was prepared by dissolving
50mg of Aceclofenac in sufficient PBS to produce 50 mL solution in a volumetric
flask.Aliquot of 10 ml diluted upto 100 ml (i.e. 100 µg/ml). From the above solution
are prepared in range of 10-90 µg/ml. Absorbances were taken on UV
spectrophotometer at 273 nm against 7.4 PBS as a blank. From these absorbance’s,
the standard curve is plotted. Standard curve equation and regression value is obtained.

D. Piroxicam
a. Standard curve in 0.1N HCl
Standard stock solution of piroxicam was prepared by dissolving 100 mg drug
in 100 ml 0.1N HCL (i.e. 1000 μg/ml) . From the stock solution 10 ml was taken and
diluted upto 100 ml (i.e. 100 in 0.1N HCL. Again 0.2, 0.4, 0.6, 0.8, 1, 1.2, 1.4, 1.6, 1.8
and 2 ml solution was taken from this solution and dilute upto 10 ml with 0.1N HCL
to get the desired concentration range (2-20 μg/ml). The absorbance of drug was
measured at λmax 333 nm on UV spectrophotometer.

5.3 Results of preformulation studies

A. Lornoxicam
Identification of drug
The highest peak was found at 376 nm. Zero order derivative UV
spectrophotometric methods were used for the analysis of lornoxicam and the highest
peak was found at 367 nm. The drug samples were prepared with 0.05 N NaOH and
scanned with UV [Fig 5.1]. The values compared with standard drug volume, which is
reported in Merck Index.

1) Organoleptic Properties
Organoleptic properties of the drug sample were found to be as given in table
below-5.1

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Table 5.1 Organoleptic properties of Lornoxicam

Organoleptic properties Results
Colour Orange to Yellow powder
Crystallinity Amorphous in nature
Taste Slightly bitter in taste
Odour Odourless

2)Partition Coefficient
Partition Coefficient of Lornoxicam was found to be 1.7. The above value of
partition coefficient is nearby to the value of partition coefficient reported in Merck
index for Lornoxicam. The partition coefficient shows that the drug is lipophilic in
nature which makes it suitable for transdermal delivery via Pluronic lecithin
organogel.

3)Melting Point
Melting Point of Lornoxicam was found in range of 225-2280c which falls
under the melting point range specified in Merck index. This shows that the drug is
pure. The drug starts melting at 225º c and completes melting at 228º c which indicates
amorphous nature of drug.

4)Solubility Properties
Qualitative Solubility: The results of qualitative solubility of the drug in different
solvents are given below in the table 5.2.

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Table 5.2 Qualitative Solubility of drug in different solvents

Solvents(5ml) Solubility Properties of the drug(5mg)
Distilled Water +

0.1N HCl +++

3.6 pH Buffer ++

7.4 pH Buffer +++

9.2 pH Buffer +++

Ethanol ++

Methanol ++

Chloroform ++

Acetone +++

Hexane +
0.05 N NaOH ++++

+ Insoluble
++ Poorly soluble
+++ Slightly soluble
++++ Freely soluble

The results were showed that Lornoxicam is insoluble in distilled water &
hexane and very less solubility in organic solvents like ethanol, methanol, chloroform
& acetone but the drug was freely soluble in alkaline solvents like 7.4 pH buffer and
9.2 pH buffer. The drug showed high solubility in 0.05 N NaOH, which indicates the
acidic nature of the drug.

Quantitative Solubility: The results of quantitative solubility of the drug are given
below in the table 5.3.

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Table 5.3 Quantitative Solubility of drug in different solvents

Solvent Concentration of drug in solvent

0.05 N NaOH 6.306 mg of drug was present in 1ml of 0.05 N NaOH

0.1N HCl 0.664 mg of drug was present in 1 ml of 0.1N HCl

3.6 pH Buffer 0.732 mg of drug was present in 1 ml of 3.6 pH buffer

7.4 pH Buffer 0.92 mg of drug was present in 1 ml of 7.4 pH buffer

9.2 pH Buffer 1.224 mg of drug was present in 1 ml of 9.2 pH buffer

The observations showed that the solubility of Lornoxicam increases with the increase
of pH from 3.0 to 9.0, which indicates that the ionization of drug increases with the
elevating pH.

5.) Standard Curve

Standard curve of the drug in 0.05 N NaOH & 7.4 PBS was prepared by
method reported by Nemutlu et al (2005). The absorbances were taken out at 376 nm.

a. Standard curve of lornoxicam in 0.05 N NaOH

Absorbance’s of the drug at 376 nm in 0.05 N NaOH are given below in table
5.4.
Table 5.4 Absorbances of Lornoxicam at 376 nm in 0.05 N NaOH

S. Concentration (µg/ml) Absorbance

No.
1 5 0.076
2 10 0.147
3 15 0.216
4 20 0.298
5 25 0.367
6 30 0.438
7 35 0.513

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Standard curve of Lornoxicam in 0.05 N NaOH at 376 nm is shown below in fig. 5.1

Fig 5.1: Standard Curve of Lornoxicam in 0.05 N NaOH

b. Standard curve of lornoxicam in PBS 7.4:

Absorbances of the drug at 376 nm in PBS 7.4 are given below in table 5.5-

Table 5.5 Absorbances of Lornoxicam at 376 nm in PBS 7.4

S. No. Concentration Absorbance
(µg/ml)

1 0 0

2 4 0.1672

3 8 0.3354

4 12 0.4885

5 16 0.6515

6 20 0.8225

7 24 0.9663

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Fig 5.2 Standard Curve of Lornoxicam in 7.4 PBS

Fig. 5.1 & 5.2 showed linearity in the range of 5-35 µg/ml & 4-24 µg/ml with
regression coefficient of 0.9997 & 0.9996 respectively. It shows that the drug follows
Beer’s Lambert Law in these ranges.

7) Particle Size
The results of the Microscopic evaluation for the measurement of particle size
of the drug particles are given below in table 5.6.

Table 5.6 Particle size distribution of Lornoxicam

S. No. Size Range Mid Point No. of Particles M. P. × M. P. × N × L. C.
(M. P.) (N) N (d)

1. 0-1 0.5 06 03 5.82

2. 1-2 1.5 09 13.5 12.15

3. 2-3 2.5 11 27.5 53.35

4. 3-4 3.5 27 94.5 183.33

5. 4-5 4.5 23 103.5 200.79

6. 5-6 5.5 24 132 256.06

∑n=100 ∑d=714.5

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Least Count (L. C.) = 1.94

Particle size was found to be 7.145 µm. Particle size distribution pattern
depicted in fig. 5.4 shows that drug particles are distributed in a range of 1-6 µm and
maximum number of particles are present in size range of 4-6 µm. This distribution
pattern also indicates that the drug is amorphous in nature.

From the above data particle size distribution graph is plotted which is shown in fig.
5.3.

Fig. 5.3 Particle Size Distribution of Drug (Lornoxicam)

8)Drug- excipient compatibility studies

Drug-excipient compatibility is performed by method described in merck
index. This study is done to find out compatibility between the drug and excipients.
From the results given in table 5.6, it can be concluded that there is no interaction
between excipients and drug. The drug and excipient are compatible with each other
and can be used for formulation of Gel.
The results of Drug-Excipient Compatibility studies are shown in table 5.7.

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Table 5.7 Drug-Excipient Compatibility Observations

S.No. Additives (50 mg each) Observation at Observation at 40°C Remarks
with drug 60°C for 2 weeks for 2 month

1. Drug ( lornoxicam) No interaction No interaction Accepted

2. Drug + pluronic F-127 No interaction No interaction Accepted
3. Drug + lecithin No interaction No interaction Accepted
4. Drug + isopropyl myristate No interaction No interaction Accepted
5. Drug + PEG 400 No interaction No interaction Accepted
6. Drug + Sodium sorbate No interaction No interaction Accepted
7. Drug + Sodium Benzoate No interaction No interaction Accepted
8. Drug + Triethanolamine No interaction No interaction Accepted
9. Drug + carbopol 934 No interaction No interaction Accepted
10. Drug + Oleic acid No interaction No interaction Accepted
11. Drug + Ethanol No interaction No interaction Accepted
12. Drug + propylene glycol No interaction No interaction Accepted

A. Flurbiprofen
Identification of drug: The λmax of Flurbiprofen was obtained by using double beam
uv visible spectrophotometer in the range of 230-360 nm..The maximum peak was
observed at 248 nm which is same as reported in I.P.
1) Organoleptic properties:
On the basis of organoleptic properties it was found that flurbiprofen was white,
crystalline powder, bitter taste and almost odorless shown in table 5.8.

Table 5.8: Organoleptic properties of flurbiprofen

S.No. Parameter Description
I) Color White Or Almost White.
II) Crystallinity Crystalline Powder.
III) Taste Bitter
IV) Odors Odorless Or Almost Odorless.

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2) Solubility determination: It was found that flurbiprofen was soluble in most of the
organic solvent and insoluble in water as shown in table 5.9.
Table 5.9 Solubility of Flurbiprofen
S.No. Parameter Description
I) Qualitative Insoluble In Water.
Freely Soluble In-Ethanol(95 %),
Methanol ,Chloroform,
Acetone, Ph 7.4 Buffer

Quantitative solubility: The results of Quantitative solubility of the drug are given
below in the following table 5.10.

Table 5.10: Quantitative Solubility of drug in different solvents

Solvent Concentration of drug in solvent
Ethanol(95%) 7.412 mg of drug was present in 1ml of ethanol
Methanol 8.365 mg of drug was present in 1 ml of Methanol
7.4 pH Buffer 6.201 mg of drug was present in 1 ml of 7.4 pH buffer
Distilled water 0.03 mg of drug was present in 1 ml of distilled water

3) Partition co-efficient: The partition coefficient of Flurbiprofen in chloroform/water

was found to be 3.89. The obtained value of partition coefficient is suitable for
transdermal drug delivery.

4) Particle Size
The results of the microscopic evaluation for the measurement of particle size of the
drug particles are given below in table 5.11.

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Table 5.11: Particle size distribution of Flurbiprofen

S. No. Size Range Mid Point No. of Particles M. P.* N M. P. * N *
(M. P.) (n) L. C.(d)
1. 0-1 0.5 06 03 5.82
2. 1-2 1.5 09 13.5 12.15
3. 2-3 2.5 11 27.5 53.35
4. 3-4 3.5 27 94.5 183.33
5. 4-5 4.5 23 103.5 200.79
6. 5-6 5.5 24 132 256.06
∑n=100 ∑d=714.5
Particle size of Flurbiprofen = ∑d/∑n
=714.5/100
=7.145µm

From the above data particle size distribution graph is plotted which is shown in fig.
5.4.

Fig. 5.4 Particle Size Distribution of Drug

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Particle size was found to be 7.145 µm. & distribution pattern depicted in fig. 5.4 .The
drug particles are distributed in a range of 1-6 µm and maximum number of particles
are present in size range of 4-6 µm.

5) Melting point:
The melting point of flurbiprofen was obtained by Thiels melting point apparatus. The
melting point was observed from 110-1120C which is approximately same as I.P.1996.

6) Standard curve in ethanol:

Standard curve of flurbiprofen in ethanol: the standard curve of flurbiprofen was
prepared in ethanol as shown in table 5.12 and fig 5.5

Table 5.12 Data for standard curve of flurbiprofen in ethanol

S.No. Concentration (μg / ml) Absorbance
1. 0 0

2. 2 0.098

3. 4 0.185

4. 6 0.368

5. 8 0.467

6. 10 0.599

7. 12 0.743

8. 14 0.879

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Fig 5.5. Standard curve of flurbiprofen in ethanol

h). Standard curve of flurbiprofen in 7.4 PBS

Standard curve of flurbiprofen in 7.4 PBS: the standard curve of flurbiprofen was
prepared in 7.4 PBS as shown in table 5.13 and fig 5.6

Table 5.13: Absorbance of Flurbiprofen at 248 nm in PBS 7.4

S. No. Concentration (µg/ml) Absorbance

1 0 0
2 10 0.149
3 20 0.248
4 30 0.350
5 40 0.451
6 50 0.562
7 60 0.628
8 70 0.738
9 80 0.837
10 90 0.902

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Fig 5.6. Standard curve of flurbiprofen in Pbs 7.4

7) Drug- Excipient compatibility studies

Table 5.14 Drug-Excipient Compatibility Observations
S.No. Additives (50 mg each) Observation at Observation at Remarks
with drug 60°C for 2 weeks 40°C for 2 month

1. Drug (flurbiprofen) No change No change Accepted

2. Drug + pluronic F-127 No change No change Accepted

3. Drug + lecithin No change No change Accepted

4. Drug + isopropyl myristate No change No change Accepted

5. Drug + PEG 400 No change No change Accepted

6. Drug + Sodium sorbate No change No change Accepted

7. Drug + Sodium Benzoate No change No change Accepted

8. Drug + Oleic acid No change No change Accepted

9. Drug + Ethanol No change No change Accepted

10. Drug + propylene glycol No change No change Accepted

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From the results given in table 5.11. it is concluded that there is no interaction between
excipients and drug. The drug and excipient are compatible with each other and can be
used for formulation of gel.

B. Aceclofenac
Identification of drug
The highest peak was found at 273 nm. The UV scan of the drug sample fig 5.7
showed highest peak at 276 nm which is nearby to the standard value reported in the
Merck index.

1)Organoleptic properties
Organoleptic properties of the drug sample was found to be as given in table
below.

Table 5.15: Organoleptic properties of Aceclofenac

Organoleptic properties Results

Colour Pale Yellow powder

Crystallinity Amorphous in nature

Taste Slightly bitter in taste

Odour Odourless

2)Partition coefficient
Partition Coefficient of Aceclofenac was found to be 1.3. The above value of
partition coefficient is nearby to the value of partition coefficient reported in Merck
index for Aceclofenac. The partition coefficient shows that the drug is lipophilic in
nature which makes it suitable for transdermal delivery via Pluronic lecithin
organogel.

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3)Melting point
Melting Point of Aceclofenac was found in range of 273-2780c which falls
under the melting point range specified in Merck index. This shows that the drug is
pure. The drug starts melting at 275º c and completes melting at 278º c which indicates
amorphous nature of drug.

4)Solubility Properties
Qualitative Solubility: The results of qualitative solubility of the drug in
different solvents are given below in the table 5.16.

Table 5.16 Qualitative Solubility of drug in different solvents

Solvents(5ml) Solubility Properties of the drug(5mg)
Distilled Water ++
0.1N HCl +++
3.6 pH Buffer +
7.4 pH Buffer ++++
9.2 pH Buffer +
Ethanol +++
Methanol ++
Chloroform ++
Acetone +
Hexane +
0.05 N NaOH ++

+ Insoluble
++ Poorly soluble
+++ Slightly soluble
++++ Freely soluble
The results showed that Aceclofenac is insoluble in acetone & hexane and very
less solubility in organic solvents like, methanol, chloroform. The drug showed high
solubility in 0.1 n HCL and ethanol, which indicates the acidic nature of the drug.

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Quantitative solubility: The results of Quantitative solubility of the drug are given
below in the table 5.17.
Table 5.17: Quantitative Solubility of drug in different solvents
Solvent Concentration of drug in solvent
0.05 N NaOH 1.324mg of drug was present in 1ml of 0.05 N NaOH
0.1N HCl 3.745 mg of drug was present in 1 ml of 0.1N HCl
3.6 pH Buffer 0.561 mg of drug was present in 1 ml of 3.6 pH buffer
7.4 pH Buffer 5.323 mg of drug was present in 1 ml of 7.4 pH buffer
9.2 pH Buffer 0.923 mg of drug was present in 1 ml of 9.2 pH buffer

5)Standard curve
Standard curve of the drug in 7.4 PBS was prepared. The absorbances were
taken out at 273 nm.
Standard curve of Aceclofenac in PBS
Absorbance’s of the drug at 273 nm in 0.05 N NaOH are given below in table 5.18

Table 5.18: Absorbances of Aceclofenac at 273 nm in PBS

S. No. Concentration (µg/ml) Absorbance
1 5 0.081
2 10 0.141
3 15 0.221
4 20 0.289
5 25 0.361
6 30 0.429
7 35 0.520

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Standard curve of Aceclofenac in PBS at 273 nm is shown below in fig. 5.7

Fig 5.7: Standard Curve of Aceclofenac in 0.05 N NaOH

Fig. 5.9 showed linearity in the range of 5-35 µg/ml & 4-24 µg/ml with
regression coefficient of 0.9993. This shows that the drug follows Beer’s Lambert Law
in these ranges.

6) Particle size
The results of the Microscopic evaluation for the measurement of particle size
of the drug particles are given below in table 5.19.

Table 5.19 Particle size distribution of Aceclofenac

S. No. Size Range Mid Point No. of Particles M. P. × M. P. × N × L. C.
(M. P.) (N) N (d)
1. 0-1 0.5 05 2.5 4.625
2. 1-2 1.5 09 13.5 24.975
3. 2-3 2.5 14 35 64.75
4. 3-4 3.5 26 91 168.35
5. 4-5 4.5 21 94.5 174.85
6. 5-6 5.5 25 137.5 254.375
∑n=100 ∑d=691.925

Least Count (L. C.) = 1.85

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Particle size was found to be 6.91 µm. Particle size distribution pattern
depicted in fig. 7.4 shows that drug particles are distributed in a range of 1-5 µm and
maximum number of particles are present in size range of 4-7 µm. This distribution
pattern also indicates that the drug is amorphous in nature.

Fig. 5.8 Particle size distribution of drug (Aceclofenac)

6)Drug- excipient compatibility studies
Drug-excipient compatibility is performed by method described in merck
index. This study is done to find out compatibility between the drug and excipients.
From the results given in table 5.20 it can be concluded that there is no interaction
between excipients and drug. The drug and excipient are compatible with each other
and can be used for formulation of Gel. The results of Drug-Excipient Compatibility
studies are shown in table 5.20.

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Table 5.20 Drug-Excipient Compatibility Observations

S.No. Additives (50 mg each) Observation at Observation at Remarks
with drug 60°C for 2 weeks 40°C for 2 month

1. Drug ( Aceclofenac) No interaction No interaction Accepted

2. Drug + pluronic F-127 No interaction No interaction Accepted

3. Drug + lecithin No interaction No interaction Accepted

4. Drug + isopropyl myristate No interaction No interaction Accepted

5. Drug + PEG 400 No interaction No interaction Accepted

6. Drug + Sodium sorbate No interaction No interaction Accepted

7. Drug + Sodium Benzoate No interaction No interaction Accepted

8. Drug + Triethanolamine No interaction No interaction Accepted

9. Drug + carbopol 934 No interaction No interaction Accepted

10. Drug + Oleic acid No interaction No interaction Accepted

11. Drug + Ethanol No interaction No interaction Accepted

12. Drug + propylene glycol No interaction No interaction Accepted

D. Piroxicam
Identification of drug: The λmax of Piroxicam was obtained by using uv visible
spectrophotometer in the range of 230-400 nm.The maximum peak was observed at
354 nm which is same as reported in I.P-1996.

1) Organoleptic properties:
On the basis of organoleptic properties it was found that Piroxicam was white
powder, bitter in taste and almost odorless shown in table 5.21.

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Table 5.21: Organoleptic properties of Piroxicam

S.No. Parameter Description

I) Color Off white to light yellow powder

II) Crystallinity Crystalline Powder.

III) Taste Sligtly Bitter in taste

IV) Odors Odorless

2) Solubility Determination Qualitative Solubility-

The value of qualitative solubility of piroxicam are shown below
Table 5.22: Qualitative solubility of piroxicam
Solvents (5 ml) Solubility properties of drug
Distilled water +
Acetone +++
Methanol +++
Ethanol ++
Simulated salivary fluid (6.8 pH) ++
0.1N HCL ++
Chloroform ++

Where-
+ : Poorly soluble, + + : Slightly soluble, + + + : soluble
Qualitative solubility studies of drug shown in table 5.22 depicted that the drug is
more soluble in organic solvents as compare to hydrophilic solvents so it can be
concluded that drug is lipophilic in nature.

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Quantitative solubility:
The result of quantitative solubility of piroxicam are given below-
Table 5.23: Quantitative Solubility of Piroxicam
Solvents (2 ml) Solubility (mg/ml)
Distilled water 0.027mg/ml
Acetone 22.4 mg/ml
Methanol 7.56 mg/ml
Ethanol 5.97 mg/ml
PBS (6.8 pH) 6.19 mg/ml
0.1N HCL 1.70 mg/ml

Chloroform 10.4 mg/ml

3) Partition co-efficient: The partition coefficient of Piroxicam in chloroform/water

was found to be 2.8-3. The obtained value of partition coefficient is suitable for
transdermal drug delivery.

4) Particle size
The results of the Microscopic evaluation for the measurement of particle size
of the drug particles are given below in table 5.24.

Table 5.24: Particle size distribution of Piroxicam

S. No. Size range Mid point No. of particles M.P.*No. of
(N) particle*L.C. = d
1 0-1 0.5 59 36.8
2 1-2 1.5 19 35.6
3 1-3 2.5 16 50
4 1-4 3.5 5 21.8
5 1-5 4.5 1 5.6
Total Ʃn = 100 Ʃd= 150.00

Particle size of Piroxicam was found 1.5 micrometer.

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From the above data particle size distribution graph is plotted which is shown in fig.
5.9

Fig. 5.9: Particle size distribution of drug

Particle size was found to be 1.5 µm. Particle size distribution pattern depicted in fig.
5.8 shows that drug particles are distributed in a range of 1-6 µm and maximum
number of particles are present in size range of 0-2 µm.

5) Melting point:
The melting point of Piroxicam was obtained by Thiels melting point apparatus. The
melting point was observed from 197-2000C which is approximately same as I.P.1996.

6) Standard curve in 0.1 N HCL

Standard curve calibration of piroxicam was prepared in 0.1N HCl at 333 nm and 354
nm on UV spectrophotometer. The linearity range was found between 2 – 20μg /ml
which obeys Lambert beers low.

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Table 5.25: Standard curve of Piroxicam

S.No. Concentration. (μg /ml) Absorbance at Absorbance at
333nm in 0.1N 354nm in
HCl. SSF(6.8)
0 0 0 0
1 2 0.133 0.073
2 4 0.249 0.135
3 6 0.368 0.214
4 8 0.510 0.281
5 10 0.630 0.349
6 12 0.755 0.417
7 14 0.876 0.484
8 16 1.024 0.549
9 18 1.159 0.622
10 20 1.250 0.686

Fig 5.10: Standard curve calibration of Piroxicam

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7) Drug-Excipient Compatibility studies

This study is done to find out compatibility between the drug and excipients. The
results of Drug-Excipient Compatibility studies are shown in table 5.26.

Table 5.26: Drug-Excipient Compatibility Observations

S.No. Additives (50 mg each) Observation at Observation at Remarks
with drug 60°C for 2 weeks 40°C for 2 month

1. Drug (Piroxicam) No change No change Accepted

2. Drug + pluronic F-127 No change No change Accepted

3. Drug + lecithin No change No change Accepted

4. Drug + isopropyl myristate No change No change Accepted

5. Drug + PEG 400 No change No change Accepted

6. Drug + Sodium sorbate No change No change Accepted

7. Drug + Sodium Benzoate No change No change Accepted

8. Drug + Oleic acid No change No change Accepted

9. Drug + Ethanol No change No change Accepted

10. Drug + propylene glycol No change No change Accepted

From the results given in table 5.26, it is concluded that there is no interaction between
excipients and drug. The drug and excipient are compatible with each other and can be
used for formulation of gel.

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